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KMID : 0360319930250030429
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Journal of Korean Cancer Research Association
1993 Volume.25 No. 3 p.429 ~ p.444
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Detection of Ras Gene Mutations in Human Cancer by PCR-SSCP
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김철민
박주인/강치덕/김선희/박영홍/정수자/정병선
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Abstract
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To clarify the effect of activation of ras oncogene upon the development of human skin cancer, detection of point mutations in c-Ki-ras-2 oncogene from 33 cases of formalin-fixed, paraffin-embedded basal cell carcinomas was carried out by
Single-Single-Strand Conformational Polymorphism Analysis of Polymerase Chain Reaction Products(PCR-SSCP).
All the specimens were primarily deparaffinized and then genomic DNA was purified partially. And exon 1 of c-Ki-ras-2 gene was amplified specifically by polymerase chain reaction (PCR) with radioisotope labelled primers, Comparison between normal
and
mutated genes was done by nondenaturing polyacrylamide gel electrophoresis of PCR products and confirmed by nucleotide sequence analysis with asymmetric PCR and direct sequencing method.
PCR products of 3 cases showed different migration patterns form others in polyacrylamide gel electrophoresis. Nucleotide sequences of c-Ki-ras-2 of them were changed from GGT to GAT in 2 cases and to AGT in 1 case at codon 12 position. The
mutations
are consistent with replacement of glycine by aspartate and serine respectively. They were heterogyzote because they had both normal and mutated alleles. These findings may conclude that mutational activations of c-Ki-ras-2 gene can take place in
human
basal cell carcinomas.
But the rate of mutations in human basal cell carcinomas was 9% and it seems to be relatively low. Additional study on the activation of other ras oncogenes, c-Ha-ras and N-ras, will clarify the effect of ras oncogenes on the development of human
skin
cancers more certainly.
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KEYWORD
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